Identification of a Novel Feruloyl Esterase by Functional Screening of a Soil Metagenomic Library

Abstract

A cosmid metagenomic library containing 1.3 × 105 clones was created from a soil sample. A novel gene (fae-xuan) encoding a feruloyl esterase was identified through functional screening. Primary sequence analysis showed that the gene consisted of 759 base pairs and encoded a protein of 252 amino acids. The gene was expressed in Escherichia coli BL21 (DE3) and the corresponding purified recombinant enzyme exhibited a molecular weight of 29 kDa. The FAE-Xuan showed high activity (40.0 U/mg) toward methyl ferulate with an optimal temperature and pH of 30 °C and 5.0, respectively. Besides methyl ferulate, FAE-Xuan can also hydrolyze methyl sinapate and methyl p-coumarate. The substrate utilization preferences and phylogenetic analysis indicated that FAE-Xuan belongs to type A FAE. FAE-Xuan was quite stable over a broad pH range from 3.0 to 10.0. The activity reduced remarkably in presence of Cu2+. FAE-Xuan can enhance the quantity of ferulic acid from de-starched wheat bran in presence of xylanase. The work presented here highlighted the effectiveness of metagenomic strategy in identifying novel FAEs with diverse properties for potential use in industrial production.

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  • 26 February 2019

    The original version of this article unfortunately contained a mistake in the image and caption of Fig. 6. The corrected version of the image and caption is shown here.

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Funding

This work was supported by the Fund for Qing Lan Project of Jiangsu Province, by the Fundamental Research Funds for the Central Universities (KYYJ201708), and by special funds of agro-product quality safety risk assessment of the Ministry of Agriculture of the People’s Republic of China (GJFP201701505).

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Correspondence to Zhihong Xin.

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Li, X., Guo, J., Hu, Y. et al. Identification of a Novel Feruloyl Esterase by Functional Screening of a Soil Metagenomic Library. Appl Biochem Biotechnol 187, 424–437 (2019). https://doi.org/10.1007/s12010-018-2832-1

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Keywords

  • Feruloyl esterase
  • Metagenomic library
  • Protein expression
  • Functional screening
  • pH stability